Chemical microbiological analysis of the water of Paliastomi Lake
On May 14–15, 2017, an expedition to conduct chemical analysis of the chemical, microbiological and seabed sediments of Paliastomi Lake was organized. We selected four analysis and sampling points near Paliastomi Lake, adjacent to Collinbari Island: 1. Rangers’ Base-1. 100 meters offshore from the ranger base; 2 Rangers’ Base-2, at 100 m, from the base; Rangers’ Base-3, at 500 m from Ranger base; and ranger base-4 near the fish processing plant, 150 m from the factory exit channel (Fig. 1) [13].
Paliastomi Lake water and seabed sampling, preservation, storage and transportation was done using ISO standard methods. The chemical properties of water organoleptic, chemical and microbiological indicators as well as seabed sediments were determined by field portable apparatus and stationary laboratory using ISO and GOST standard methods [15, 116].
The pH value of Paliastomi Lake water is slightly more than pronounced due to the nature of the salts dissolved in it and the hydrolysis. According to samples 1, 2 and 3, the oxygen content is high at 7.3–7.8 mg / L, and the amount of oxygen dissolved according to the 4th data is relatively low and equals to 3.6 mg / L. The oxygen regime has a profound effect on the life-ability of the reservoir, the minimum dissolved oxygen content that ensures normal fish development is 5 mg / l O2 / L, and its reduction to 2 mg / L results in massive fish mortality [5, 9]. The concentration of dissolved oxygen in samples 1, 2 and 3 corresponds to class II-purified class of surface water quality, and class 4 is classified as contaminated, class 4.
The composition of waterbed sediments in the Lake basin is regularly altered by constructing rocks in accordance with lithology and grain size [17, 18]. In addition, as a result of sedimentation of minerals and organic matter in the Lake water, the water flows into the Lake produces sediment on its bed. The heavy metal contamination of seabed sediments is mainly caused by anthropogenic sources, residues of agriculture and enterprises working in various industries [17, 18].
Paliastomi Lake sediments were sampled and studied (by ISO methods) in May of 2017. Based on the geochemical background, the results of the analysis, the total copper content in the bottom sediments of Paliastomi Lake is lower than the permissible rates [2, 15, 19, 20], and exceeds the permissible values in Georgia [2, 18]. Lead and zinc content in Lake sediments may be considered as a natural background [2, 18]. The content of manganese and iron in the seabed is less than the MPC.
Tests of surface water and Lake bed sediments
The bicrobiological analysis determined the total coliforms and the number of coli-forming bacteria in the intestinal tract. In samples 1, 2 and 4, the total coliforms are sufficiently high and in 100 ml of water they are 3000, 1280, 5000 CFU, while in the third sample, their number is 920 CFU / 100 ml respectively. E-coli-grade bacteria were found in water samples 2 and 4, with a mean value of 100 − 40 ml per 100 ml 20–40 CFU (See Table 2). Their numbers increase with the migration of migratory birds in October and May.The total water content of the open water reservoirs - mesophilic aerobes and facultative anaerobic microorganisms (MAFA) as per the permitted norm in 1 ml is no more than 1000 CFU, while in 1, 2 and 3 water samples the total amount of MAFA varies from 800 to 5000 CFU / ml, and the maximum value (3500 CFU / ml) is reached in the 4th sample (See Table 1).
Table 1
Microbiological analysis of Paliastomi Lake water (August 10, 2017)
Microbial quality indicators of Paliastomi Lake water:
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Used Standard: ГОСТ N 18963-73
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Site: N1 R. Kaparchina-Adjacent territory to the Bridge
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Microbial quality indicators of the site water
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Outdoor reservoir water quality index MAFA actual rate by ND. Not more then
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1000 CFU/ml
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Total quality of MAFA/ml, MAFA actual rate
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5000 CFU/ml
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Indicator of water quality (coli-titer; coli-index ) by ND
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coli-titer < 0,4; not less − 111, coli-index; not more-9
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Actual indicator of the quality of open water reservoir: coli-titer and coli-index: 100 ml,10 ml,1 ml,0,1 ml (Total volume of water 111 ml);
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coli-titer < 0,4
coli-index > 2380
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pH
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6,6
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Site: N 2 Rangers Base right site
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Microbial quality indicators of the site water
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Outdoor reservoir water quality index MAFA actual rate by ND. Not more then
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1000 CFU/ml
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Total quality of MAFA/ml, MAFA actual rate
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1000 CFU/ml
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Indicator of water quality (coli-titer; coli-index ) by ND
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coli-titer not less − 111, coli-index; not more − 9
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Actual indicator of the quality of open water reservoir: coli-titer and coli-index: 100 ml,10 ml,1 ml,0,1 ml (Total volume of water 111 ml);
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coli-titer − 4
coli-index − 230
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pH
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6,5
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Site: N 3Rangers Base left site
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Microbial quality indicators of the site water:
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Outdoor reservoir water quality index MAFA actual rate by ND. Not more then
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1000 CFU/ml
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Total quality of MAFA/ml, MAFA actual rate
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1000 CFU/ml
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Indicator of water quality (coli-titer; coli-index ) by ND
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coli-titer not less 111, coli-index; not more − 9
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Actual indicator of the quality of open water reservoir: coli-titer and coli-index: 100 ml,10 ml,1 ml,0,1 ml (Total volume of water 111 ml);
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coli-titer-4
coli-index − 230
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pH
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6,5
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Site: Adjacent territory of the Island Kolimbari
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Microbial quality indicators of the site water:
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Outdoor reservoir water quality index MAFA actual rate by ND. Not more then
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1000 CFU/ml
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Total quality of MAFA/ml, MAFA actual rate
|
|
Indicator of water quality (coli-titer; coli-index ) by ND
|
1000 CFU/ml
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Actual indicator of the quality of open water reservoir: coli-titer and coli-index: 100 ml,10 ml,1 ml,0,1 ml (Total volume of water 111 ml);
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Coli-titer not less − 111, coli-index; not more − 9
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Outdoor reservoir water quality index MAFA actual rate by ND. Not more then
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coli-titer- 0,4
Coli-index– 2380
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pH
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6,8
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The open water reservoir water quality index (coli titer, coli index) shall be as follows: coli -titer not less than 111 ml and coli-index not more than 9.
Coli index in water samples 1 and 4, at pH = 6.6 and pH = 6.8, was more than 2380 and coli index was less than 0.4. In the second and third samples, at pH = 6.5, the coli titre was reduced to 4 and the coli index was 230 (See Table 2). Therefore, high levels of bacterial contamination of the Lake water should be caused not only by the contamination of Paliastom Lake, but also by sewage discharges (directly into the Lake, without any treatment) from internally displaced persons’ shelter and open landfills adjacent to it.
In January 29 2018 has been taken water sample from the lake Paliastomi in 50 meters right from Rangers’s base (latitude 42.11842; longitude 41.70293) for the purification by bacteriophages.
The microbiological analyses shown that total amount of MAFAN in the water constituted 1500 CFU/ml when the permissible norms is not more than 1000 CFU/ml.
In an open water reservoir quality indicator (coli-titer and coli-index) MAC is following: coli-titer-111, coli-index-9. During the condition when pH = 6,8 coli-titer − 4 and coli-index was 230. Total amount of E-coli constituted 1x 103, but MAFAN equaled 1500 CFU/ml (See Table 3).
We used water sample as a model pattern for the bacterial treatment with bacteriophage cocktail (BC-1) (Shigella flexneri 1, 2, 3, 4, 6 serological group; Shigella sonnei; Salmonella paratyphi A; Salmonella paratyphi B; Salmonella typhimurium; Salmonella choleraesuis; Salmonella oranienburg; Salmonella enteritidis;; enteropathogenic E. coli different serological groups; Proteus; Enterococcus; Staphylococcus; Pseudomonas aeruginosa).
Table 2 Microbiological analysis of Paliastomi Lake water (29March, 18April, 2018)
The BC-1 contained viruses of the bacteria (bacteriophages) that directly affect their host bacteria, so the BC-1 was used to treat Lake Paliastom water, reducing the amount of E-coli to 2 x102 CFU/ml( See figure 2). The amount of MAFAN/ml reduced to 600 CFU / ml. We considered the experiment successful, as during the processing of the lake water sample its bacterial contamination was reduced by 60% (See Table 3).
For in-depth study of bacteriophage treatment technology for microbiological contamination of Lake Paliastomi water, samples were taken at the following facilities:
- March 21, 2019 Paliastom Lake water samples 50 meters to the right of the Ranger base;
- In the vicinity of the fish factory near the bridge.
The microbiological quality of the water was assessed according to the requirements for standard drinking water and Category 2 surface water.
Examined water according to general microbial parameters gives the typical picture of natural open water reservoirs with moderate degree of pollution. Relatively high rates of total microbial quantity (220C) and fecal contamination (by total coliforms). No coliform content was observed. However high content of vibrios and aeromonas was detected. See the results of the study in the table. 2–2-isolates of fecal coliforms, enterococci, vibrios, and salmonellae were removed (Table 4).
According to the Table 4, the general microbial parameters, the test water gives the characteristic picture of natural open water reservoirs (with small - moderate pollution). The rates of total microbial count (220C) and fecal contamination (namely total coliforms) are relatively high, although coliform content was not detected. The number of salmonella is relatively high. 2–2 isolates of fecal coliforms, enterococci, vibrios, and salmonella were sowed. The reason of low bacterial pollution might have been caused by 2 reasons: the fish factory was closed and it was not the period of waterfowl migration.
BC-1 causes lysis of following bacteria: Shigella flexneri 1, 2, 3, 4, 6 serological group; Shigella sonnei; Salmonella paratyphi A; Salmonella paratyphi B; Salmonella typhimurium; Salmonella choleraesuis; Salmonella oranienburg; Salmonella enteritidis;; enteropathogenic E. coli different serological groups; Proteus; Enterococcus; Staphylococcus; Pseudomonas aeruginosa;
BC-2 causes lysis of following bacteria: Streptococcus, Staphilococcus, E.Coli, Pseudomanas aeruginosa, Proteus;
BC-3 causes lysis of following bacteria: S.aureus, S.epidermidis, S.pyogenes, S.sanguis, S.salivarius, S.agalactiae;
BC-4 causes lysis of following bacteria: S.aureus;
BC-5 causes lysis of following bacteria: Streptococcus, Staphyloccccus, Escherichia coli, Pseudomonas aeruginosa, Proteus ;
BC-6 causes lysis of following bacteria: Shigella flexneri 1.2.3.4 and Shigella zonnei, S.typhimurium, S.enteritidis, S.newport, S.heidelberg, S.cholera suis S.oranienburg, S.dublin, S.anatum;
BC-7 causes lysis of following bacteria: S.aureus da S.epidermidis, S.pyogenes, S.sanguis, S.salivarius, S.agalactiae, E.coli.
In 30.08 2004 instruction on preventive- treatment and water purification by phages preparations against salmonella bacteria was approved by the State Veterinary Department of Georgia.
The BC-1 was tested within the WB Georgian Agriculture Development Program’s Project implemented by ICFER- CGS 03–70 “Demonstration and application of achievement of environmentally friendly and efficient technology on the use of treatment-preventive veterinary biopreparation of bacteriophages against antibiotic resistant salmonella infections in poultry.”
Chicken is given BC-1 a preparation with water and food as a prophylactic medicine. As a precaution, 10 milliliters of drinking water are mixed with 10 milliliters of BC-1 before being given. Birds are fed 2 milliliters in 10 milliliters of water for treatment purposes. For preventive measures, 1 milliliter of food-safe preparation is mixed with 1 milliliter water and sprayed on 20 grams of food. In terms of treatment, 1 milliliter of BC-1 is combined into 10 milliliters of food. The food is vigorously churned and fed to the birds to ensure an even distribution of preparation in the food. The chicken is given the preparation once a day in the morning. The preventive course lasts about 4–5 days. The treatment lasts 7–8 days. The average –preventive daily dose for a 0-60-day-old chicken is 2X106, or 4X106 for 180-day-old chickens, and 8X106 for chickens older than 180 days. In the event of an epidemic, however, the use of antibiotics is not prohibited.
We suggest to adapt the proven methodology to wild birds, and do not exclude the possibility of conducting a study to apply the methodology in aquaculture in the marine of freshwater cages, however, in this case, we advocate that fish food should be processed with a specially compiled bacteriophage cocktail, which, if necessary, would not exclude the complex use of bacteriophages with antibiotics, which will significantly reduce the proportion of their use in aquaculture.
Water Pollution Modeling
The dissemination of pathogenic bacteria in the lake water was modeled. The following data is required to develop this model system:
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Hydrodynamic characteristics of the water depths;
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Water level;
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Wind velocity;
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Bacterial contamination points;
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Bacterial monitoring;
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Number of common coliforms.
For computational modeling of metals and coliform dissemination in Lake Paliastomi, a two-dimensional numerical model based on the system of integration by vertical integration of water motion and continuity equations was used [20, 21]. The program was compiled in Visual C ++. The model is based on the integration of the two-dimensional flow dynamics of the Lake and passive impedance in the water area using a modern high-resolution numerical scheme. In the integration process, the method of visualizing the results is applied.
The occurrence of currents in Paliastomi Lake and distribution of pollutants during the northeast wind were discussed.
The real relief of Paliastom Lake was used in the modeling process, to determine: the vector fields of the lake currents, the free surface of the water, and the concentration of impurities in the lake water area.
On a regular grid (size 26 x 29), experimental data were recorded. The data of the common coliforms obtained experimentally was transferred using linear interpolation by weights. The isolines on the drawing represent each value.
In the case of the Northeast wind, numerical (simulation) experiments were performed using the data of microbiological analysis obtained by us and the above-mentioned computer model, and the distribution of coliforms in Paliastom Lake was obtained. The results are presented in Figs. 3 and 4.
Figure 3 of May 15, 2017 shows the isoforms of the distribution of coliforms on the surface of the Paliastomi lake obtained by modeling. The map shows that the maximum concentration of coliforms is obtained in the south-eastern part of the lake where 3 sources of pollution are located at Ranger Base. They are 100 meters (left and right) and 500 meters away from the base of the Rangers Base (See Fig. 3).
Figure 4 shows the distribution of computer-modeled coliforms in the case of 2 contaminants on March 29, 2018 (airport canal and confluence of the Phichora River with the Paliastom). The obtained distribution is significantly different from the images given in Fig. 3. The maximum concentrations are located at 2 points in Lake Paliastomi (north-west and north-east). The horizontal concentration gradients are approximately the same and decrease from north-east to north-west and south, which is due to meteorological conditions caused by local water circulation and turbulent diffusion formed (See Fig. 4).
The Fig. 4 model was used to determine the change in the values of the total coliforms identified by the experiment between May 15, 2017 and March 29, 2018 - April 18, 2018.
Has been developed models to show the change in concentrations of total coliforms over 24 hours. There are 2 processes - diffuse and advectional transfer of common coliforms, resulting in a decrease in concentration in the maximum zone and transfer of common coliforms in the north-east direction, which coincides with the flow formed in Lake Paliastomi. The cyclonic current formed in the eastern part of the lake causes dissemination of common coliforms in the north-eastern part of the lake.
The currents formed in Lake Paliastomi cause a change in the distribution of total coliforms, which is characterized by a decrease in concentration of 40 CFU / ml over 24 hours near the confluence of the Phichora River with Lake Paliastomi, as well as an increase of 32 CFU / ml in the vicinity of the fish farm.
We can conclude that the circulatory processes caused by turbulence in Lake Paliastomi lead to the redistribution of common coliforms throughout the water area. The distribution is uneven and especially intense in the middle of the north-western shore of the lake.
The given technology allows similar calculations to be performed for any reservoir with shallow depth. To do this, the user will only need to change the characteristics of the reservoir (horizontal dimensions, seabed relief and atmospheric wind).
The results thus obtained show that the general picture of microbiological contamination of Paliastom Lake is heterogeneous. It depends on the location of the sources of pollution, the intensity of pollution and the water currents of the lake formed by meteorological conditions.
The model proposed in the paper describes qualitatively and quantitatively well the physical process taking place in a real environment and it can be used to predict changes in the dynamic characteristics of the relevant type of reservoirs. Through this model it is possible to further solve tasks such as the rational use of environmental resources and predict the ecological status [26].